Analysis of genetic variability in human respiratory syncytial virus by the RNase A mismatch cleavage method: subtype divergence and heterogeneity.
We have applied the RNase A mismatch cleavage method to the analysis of genetic variability among human Respiratory Syncytial (RS) viruses. Antisense RNA probes of the Long strain were hybridized to total RNA extracted from cells infected with other strains. The RNA:RNA heteroduplexes were digested with RNase A and the resistant products analyzed by gel electrophoresis. Each virus generated characteristic band patterns with the different probes. Comparative analyses of the cleavage patterns indicate that antigenic subtypes correlate with genetically distinct viral groups. Viruses within each subtype, however, show substantial genetic heterogeneity and progressive accumulation of genetic changes with time. This heterogeneity is also observed among viruses of the same epidemic outbreak which cannot be distinguished with a panel of monoclonal antibodies. Different genes and gene regions also differ in their rates of change. These results are discussed in terms of RS virus evolution.
Cristina
J
J
Servicio de Biología Molecular, Centro Nacional de Microbiología, Madrid, Spain.
López
J A
JA
Albó
C
C
García-Barreno
B
B
García
J
J
Melero
J A
JA
Portela
A
A
D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D014644:Genetic Variation; D006801:Humans; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015347:RNA Probes; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D012136:Respiratory Syncytial Viruses / Q000235:genetics*; D015183:Restriction Mapping; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid
D004279:DNA, Viral; D015347:RNA Probes; D012367:RNA, Viral; D012259:Ribonuclease, Pancreatic